摘要
目的观察探讨糖尿病（DM）大鼠视网膜组织中脯氨酰羟化酶（PHI）12）的表达及意义。方法雄性Wistar大鼠108只，随机分为DM模型组和正常对照组，分别为60、48只。对DM模型组大鼠进行一次性腹腔注射1％链脲霉素枸橼酸（STZ）溶液建模，正常对照组腹腔注射等量的枸橼酸缓冲液。造模后1、3、6个月，荧光显微镜下观察大鼠视网膜血管分布形态。造模后1～6个月，采用伊凡思蓝（EB）灌注视网膜铺片检测视网膜组织内EB浓度；免疫组织化学染色法观察大鼠视网膜PHD-2阳性染色的分布特点；蛋白质免疫印迹法（Western blot）检测大鼠视网膜中PHD-2、缺氧诱导因子-1α（HIF-1α）及血管内皮生长因子（VEGF）的表达。结果荧光显微镜观察结果显示，正常对照组大鼠视网膜血管走行规则，浅深层血管形态清晰；DM模型组大鼠视网膜血管渗漏持续存在。造模后1～6个月，DM模型组大鼠视网膜血管外组织内EB含量较正常对照组明显增加，差异有统计学意义（t=3．810，2．722，2．845，2．342，2．456，3．823；P〈0．05）。免疫组织化学染色结果显示，DM模型组及正常对照组大鼠视网膜中均可见PHD-2阳性染色，主要分布于视网膜内核层、神经节细胞层及血管壁。Westernblot检测结果显示，DM模型组大鼠视网膜中PHD-2表达在造模后1、2个月时较正常对照组下降（t=16．230，16．390；P〈0．05）；HIF-1α表达在造模后1、2、3个月时较正常对照组明显升高（t=27．073，36．709，10．176；P〈0．05）；VEGF表达在造模后1～6个月均较正常对照组升高（t=13．547，31．984，21．897，8．912，9．019，14．046；P〈0．05）。结论PHD-2在DM大鼠视网膜组织中有丰富表达。PHD-2可能在糖尿病视网膜病变发生发展中有一定的调控作用，其作用途径及机制与VEGF作用通路有关。
Objective To investigate the expression and prolyl hydroxylase （PHD）-2 in retina of diabetic rats. Methods Wistar rats were randomly divided into the control group （n=48） and the diabetes group （n= 60）. The rats in diabetes group were induced with streptozotoein （STZ） injection creating a diabetic retinopathy model. The same volume of citric acid buffer was injected into the rats in the control group. Fluorescence microscope was used to observe the retinal vasculature at one, three and six months after injection. Evans blue perfusion was used to detect the blood-retinal barrier （BRB） permeability. Immunohistochemical staining was used to observe the distribution of PHD-2 positive staining. Western blot was used to measure the protein expression of PHD-2, hypoxia inducible factor（HIF）-la and vascular endothelial growth factor （VEGF） every month from one to six months after injection. Results The vascularization was normal and form was clear in retina of rats in control group. The retinal blood vessels of rats in diabetes group showed significantly increased fluorescence. Compared with the control group, the BRB permeability was significantly increased in diabetes group （P〈0.05）. Abundant expression of PHD-2 protein was detected in the inner layers of retina in control and diabetes group. Compared with the control group, the PHD-2 expression was decreased in diabetes group at one and two months after injection （t= 16. 230, 16. 390;P〈0.05）. The HIF-1α expression was significantly increased in diabetes group at one, two and three months after injection （t=27. 073, 36. 709, 10. 176; P〈0.05）. The VEGF expression was significantly increased in diabetes group every month from one to six months after injection （t = 13. 547, 31. 984, 21. 897, 8. 912, 9. 019, 14. 046; P〈0.05）. Conclusions There is abundant expression of PHD-2 in the inner layer of retina in diabetic rats. PHD-2 may play an important role in diabetic retinopathy, which is correlated with VEGF.
出处
《中华眼底病杂志》
CAS
CSCD
北大核心
2012年第3期264-267,共4页
Chinese Journal of Ocular Fundus Diseases
基金
广东省科技计划项目（20098060700106）
广州市科技计划项目（11C22060787）
